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Team:Bielefeld-CeBiTec/Notebook/Journal/CO2-fixation/Sep
From 2014.igem.org
(Difference between revisions)
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<ul> <!-- csoS1-4 Fusion mit can --> | <ul> <!-- csoS1-4 Fusion mit can --> | ||
| - | + | <li><b><i>can</i> and <i>csoS1-4</i></b></li> | |
<ul> | <ul> | ||
<li>This week we tried to find positive clones of our transformation.</li> | <li>This week we tried to find positive clones of our transformation.</li> | ||
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<li><a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Protocols#PurificationPromega" target="_blank">Plasmid isolation</a> of pSB1C3_can_csoS1-4</li> | <li><a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Protocols#PurificationPromega" target="_blank">Plasmid isolation</a> of pSB1C3_can_csoS1-4</li> | ||
</ul> | </ul> | ||
| + | <ul> | ||
| + | <li><a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Protocols#RestrictionDigestion" target="_blank">Restriction digestion</a> with <a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/KitsAndEnzymes#RestrictionEnzymes" target="_blank"><i>EcoR</i>I</a> and <a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/KitsAndEnzymes#RestrictionEnzymes" target="_blank"><i>Pst</i>I</a> </li> | ||
| + | <ul> | ||
| + | <li>Bands as expected (~2100 bp and ~3300 bp)</li> | ||
| + | </ul> | ||
| + | </ul> | ||
| + | |||
| + | </ul> | ||
| + | </ul> | ||
<br> | <br> | ||
| - | + | <ul> | |
| + | <li><b><i>Hneap</i> and <i>T7</i></b></li> | ||
<ul> | <ul> | ||
<li>This week we tried to assemble the <i>T7</i> promotor with the <i>Hneap</i>.</li> | <li>This week we tried to assemble the <i>T7</i> promotor with the <i>Hneap</i>.</li> | ||
Revision as of 10:10, 28 September 2014
 |
September |
 |
- csoS1-4
- We tried to amplify csoS1-4 (shell proteins) for a fusion with GFP.
- PCR amplification (fw-GFP-csoS1B, rv-rv-GFP-csoS1A)
- Annealing temperature: 54 °C
- Bands as expected (~4000 bp (~1800 bp csoS1-4, ~2200 bp pSB1C3))
- PCR products were purified
- csoS1-4 and GFP
- We tried to assemble the shell proteins of the carboxysome and GFP.
- Gibson Assembly with pSB1C3_csoS1-4 and GFP
- Transformation with electrocompotetent cells
- can and csoS1-4
- This week we tried to find positive clones of our transformation.
- Colony PCR (VF-Primer, VR-Primer)
- Annealing temperature: 55 °C
- Bands as expected (~3600 bp)
- Plasmid isolation of pSB1C3_can_csoS1-4
- Restriction digestion with EcoRI and PstI
- Bands as expected (~2100 bp and ~3300 bp)
- Hneap and T7
- This week we tried to assemble the T7 promotor with the Hneap.
- BioBrick Assembly (Suffix)
- Transformation with electrocompotetent cells
- Colony PCR (VF-Primer, VR-Primer)
- Annealing temperature: 55 °C
- Bands as expected (~2300 bp)
